Mechanisms and regulation of extracellular vesicle traffic in the nervous system

神经系统细胞外囊泡运输的机制和调节

• 批准号: 10063578
• 负责人: Avital Adah Rodal
• 金额: $ 35.38万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063578
• 关键词: Acute; Address; Alzheimer' s Disease; Animals; Autophagocytosis; Behavior; Biogenesis; Biological; Biology; Brain; Cells; Cellular Membrane; Chemicals; Communication; Complement; Complex; Coupled; Cultured Cells; Cytoplasm; Data; Degradation Pathway; Detection; Disease; Drosophila genus; Electron Microscopy; Equilibrium; Foundations; Gene Expression; Gene Expression Regulation; Genetic; Genetic Transcription; Goals; Guanosine Triphosphate Phosphohydrolases; Health; Human; Image; Imaging Techniques; Label; Lighting; Measurement; Measures; Mediating; Membrane; Membrane Protein Traffic; Methods; Microscopy; Modeling; Molecular; Nature; Nervous System Physiology; Nervous system structure; Neurodegenerative Disorders; Neurologic; Neuromuscular Junction; Neurons; Parkinson Disease; Pathologic; Physiological; Physiology; Presynaptic Terminals; Proteins; Recycling; Regulation; Research; Resolution; Rest; Role; Signal Transduction; Sorting - Cell Movement; Structure; Synapses; Synaptic Membranes; Synaptic plasticity; System; Testing; Tissues; Vesicle; cell type; experimental study; extracellular vesicles; in vivo; insight; live cell imaging; nervous system disorder; novel; presynaptic; reconstitution; response; targeted treatment; tool; trafficking; uptake; vesicular release

The goal of this project is to understand the molecular mechanisms of extracellular vesicle (EV) trafficking in the nervous system in vivo. EVs are small vesicles secreted from donor cells that can carry a diverse array of cargoes, and have recently come to the forefront as a novel mode of intercellular traffic and communication in the brain. EVs are thought to contribute to many human health conditions, including the spread of pathological proteins in neurodegenerative disease. However, because most studies of EVs are conducted with cells in culture, little is known about the release, uptake and fate of EVs in the diverse interacting cell types of the intact nervous system. We have developed a system in which to study traffic of endogenous neuronal EV cargoes in a living animal, using cutting edge genetic and cell biological tools available in Drosophila. Using this system, we discovered an unexpected role for synaptic periactive zone (PAZ) membrane remodeling machinery in EV cargo sorting, stability and release from axon terminals at the Drosophila larval neuromuscular junction. We also found that endogenous EV cargo release is dynamically regulated by neuronal activity. Finally, we made the surprising discovery that released EVs are partially protected from the target cell cytoplasm, suggesting the possibility that additional regulatory steps may govern their exposure and/or degradation upon internalization to recipient cells. Our proposed research will elucidate how cellular membrane traffic machinery controls the release, uptake, and fate of EV cargoes. To achieve these goals we will use advanced Drosophila genetics, live cell imaging techniques, structured illumination microscopy, electron microscopy, and tissue-specific detection and manipulation of EV cargoes in donor and recipient cells. Specifically, we propose: 1) To elucidate in vivo mechanisms of EV traffic and release by PAZ proteins. 2) to determine how activity regulates neuronal traffic of endogenous EV cargoes and 3) to determine the fate of NMJ EV cargoes, at rest and in response to neuronal activity. Given the conserved nature of synaptic membrane trafficking machinery, our findings and tools will lay the foundation for new insights into endogenous EV traffic in many aspects of nervous system function, including in human neurological disease.

本研究的目的是了解细胞外囊泡（EV）运输的分子机制， 体内的神经系统EV是从供体细胞分泌的小囊泡，其可以携带多种多样的免疫球蛋白。 货物，最近已经成为细胞间交通和通信的新模式， 大脑电动汽车被认为有助于许多人类健康状况，包括病理性传播， 神经退行性疾病中的蛋白质。然而，由于大多数EV的研究都是用细胞进行的， 在培养中，关于EV在不同相互作用的细胞类型中的释放、摄取和命运知之甚少。 完整的神经系统我们已经开发了一个系统，在其中研究交通的内源性神经元EV 货物在活的动物，使用尖端的遗传和细胞生物学工具，可在果蝇。使用 在这个系统中，我们发现了突触周围活动区（PAZ）膜重塑的意想不到的作用 果蝇幼虫中EV货物分选、稳定性和从轴突末端释放的机制 神经肌肉接头我们还发现，内源性EV货物释放是动态调节的， 神经元活动最后，我们有了一个令人惊讶的发现，即发布的电动汽车部分受到保护， 靶细胞的细胞质，这表明额外的调节步骤可能会控制其暴露的可能性 和/或在内化到受体细胞后降解。我们提出的研究将阐明细胞如何 膜交通机械控制EV货物的释放、摄取和命运。为了实现这些目标，我们 将使用先进的果蝇遗传学，活细胞成像技术，结构照明显微镜， 电子显微镜，以及供体和受体细胞中EV货物的组织特异性检测和操作。 具体而言，我们提出：1）阐明EV运输和释放PAZ蛋白的体内机制。2)到 确定活动如何调节内源性EV货物的神经元交通和3）确定 NMJ EV货物，在休息和响应神经元活动。鉴于突触的保守性， 膜运输机制，我们的发现和工具将奠定基础，新的见解，内源性 EV交通在神经系统功能的许多方面，包括在人类神经系统疾病。